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 Wh

 
An 
  is a protein used by the immune system to identify and neutralize
foreign objects like bacteria and viruses. Each antibody recognizes a specific
antigen unique to its target.

 

 
 are antibodies that are derived from different cell lines.

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 (
)) are antibodies that are identical because they were
produced by one type of immune cell, all clones of a single parent cell.

 

According to differences in their heavy chain constant domains, immunoglobulins
are grouped into five classes, or isotypes: ¢
,

, ¢ ,
, ¢ ,
, ¢ ,
, and ¢ .
.

IgG: IgG1 (66%), IgG2 (23%), IgG3 (7%) and IgG4 (4%) , blood and tissue liquid.
IgA:IgA1
IgA: IgA1 (90%) and IgA2 (10%), stomach and intestines
IgM: normally pentamer, ocassionally hexamer, multiple immunoglobins linked with
disulfide bonds
IgD:1%
IgD: 1% of proteins in the plasma membranes of B
B--lymphocytes, function unknown
IgE: on the surface of plasma membrane of mast cells, play a role in immediate
hypersensitive and denfensive for parasite
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 All current therapeutic antibodies are of the
IgG class.
 When the objective of antibody therapy is to
directly kill the target cell, the isotype of
choice is IgG1, since this isotype is optimal for
complement fixation.
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 6ab = 6ragment, antigen binding
 6c = 6ragment, crystalline
 The 6c fragment specifies biological properties
of the molecule, such as complement fixation,
placental transfer, skin fixation and catabolic
rate.
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 In 1975, Georges Kohler and Cesar
Milstein first fused lymphocytes to produce a
cell line which was both immortal and a
producer of specific antibodies. The two
scientists were awarded the Nobel Prize for
Medicine in 1984 for the development of this
"hybridoma The value of hybridomas to the
field was not truly appreciated until about 1987,
when MAbs were regularly produced in rodents
for diagnostics.
 
 

 
 






 

 
 
  



 


 


  



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- B lymphocytes can mutate into tumor

cells that result in a type of cancer termed
myeloma.
- Myeloma cells become ³immortal´ and
will grow indefinitely in culture.
- 6usion of a single activated B cell and a
myeloma cell will create a hybridoma
that can grow indefinitely in culture.
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‰ Myeloma cells have been genetically

engineered such that they can not use
hypoxanthine, aminopterin, and thymidine
(HAT medium) as a source for nucleic acid
biosynthesis and will die in culture.
‰ Only B cells that have fused with the
engineered myeloma cells will survive in
culture when grown in HAT medium.
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All the cells secrete only one type of antibody directed
against one epitope

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Practical steps in monoclonal antibody production:

1) Immunize animal

2) Isolate spleen cells (containing antibody-producing B cells)

3) 6use spleen cells with myeloma cells (polyethylene glycol, virus, by electroporation)

4) Allow unfused B cells to die

5) Add aminopterin to culture to kill unfused myeloma cells

6) Clone remaining cells (place 1 cell/well and allow each cell to grow into a clone of
cells)

7) Screen supernatant of each clone for presence of the desired antibody.

8) Grow the chosen clone of cells in tissue culture indefinitely.

9) Harvest antibody from the culture supernatant.

10) (If you¶re a biotech company) charge about $1,000-$2,000 per mg.
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‰ Mouse monoclonal antibodies have been

genetically engineered to replace all of the
antibody molecule with human counterparts
except the hyper variable regions directly
involved with antigen binding.
‰ Humanized monoclonal antibodies are
currently be tested in human clinical trials.
USES

|easuring protein and drug levels in serum

]yping tissue and blood
dentifying infectious agents
dentifying clusters of differentiation for the
classification and follow-up therapy of
leukemias and lymphomas
dentifying tumor metastasis
dentifying and quantifying hormones
mmunoaffinity Purification
 Used in several diagnostic tests to detect small amounts
of drugs, toxins or hormones, e.g. monoclonal antibodies
to HCG used in pregnancy test kits or diagnosis of AIDS
by the ELISA test.
 monoclonal antibody can be coupled to another molecule
like a fluorescent molecule to aid in imaging the target
 Or with a strongly-
strongly-radioactive atom, such as Iodine-
Iodine-131
to aid in killing the target.
 Used in the radioimmunodetection and
radioimmunotherapy of cancer, and some new methods
can even target only cancerous cells.
 Monoclonal antibodies can be used to treat viral diseases,
traditionally considered "untreatable". In fact, there is
some evidence to suggest that antibodies may lead to a
cure for AIDS.
 Monoclonal antibodies can be used to classify strains of a
single pathogen, e.g. Ú
 
   can be typed
using mAB.
 To identify and to trace specific cells or molecules in an
organism.
 OKT3, an antibody to the T3 antigen of T cells, is used to
alleviate the problem of organ rejection in patients who
have had organ transplants.
 
 

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Three mechanisms that could be responsible for the cancer treatment.

 mAbs act directly when binding to a cancer specific antigens and

induce immunological response to cancer cells. Such as inducing
cancer cell apoptosis, inhibiting growth, or interfering with a key
function.

 mAbs can be modified for delivery of a toxin

toxin,, radioisotope
radioisotope,,
cytokine or other active conjugates.

 it is also possible to design bispecific antibodies that can bind

with their 6ab regions both to target antigen and to a conjugate or
effector cell
 Herceptin (Trastuzumab) is an anti-
anti-cancer antibody
that acts on HER2/neu (erbB2) receptor, which is
overexpressed in breast cancer. Only cells
overexpressing this receptor are susceptible.

 Gemtuzumab ozogamicin (Mylotarg) monoclonal

antibody attacks the CD33 receptor, which is found in
most leukemic blast cells, but not in normal
hematopoietic stem cells
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